Quinazoline derivative

ABSTRACT

The invention concerns derivatives of the formula wherein n is 1, 2 or 3 and each R 2  is independently halogeno; and R 1  is amino-(2-4C)alkylamino, (1-4C)alkylamino, (1-4C)alkylamino-(2-4C)alkylamino,di-[(1-4C)alkyl]amino-(2-4C) alkylamino, pyrrolidin-1-yl-(2-4C)alkylamino, piperidino-(2-4C)alkylamino, morpholino-(2-4C)alkylamino, piperazin-1-yl-(2-4C)alkylamino, or 4-(1-4C) alkylpiperazin-1-yl-(2-4C)alkylamino, or pharmaceutically acceptable salts thereof; processes for their preparation, pharmaceutical compositions containing them, and the use of the receptor tyrosine kinase inhibitory properties of the compounds in the treatment of proliferative disease such as cancer.

This application is the national phase of international applicationPCT/GB96/00959, filed Apr.23, 1996 which designated the U.S.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to quinazoline derivatives, orpharmaceutically-acceptable salts thereof, which possessanti-proliferative activity such as anti-cancer activity and areaccordingly useful in methods of treatment of the human or animal body.The invention also relates to processes for the manufacture of saidquinazoline derivatives, to pharmaceutical compositions containing themand to their use in the manufacture of medicaments of use in theproduction of an anti-proliferative effect in a warm-blooded animal suchas man.

2. Description of the Related Art

Many of the current treatment regimes for cell proliferation diseasessuch as psoriasis and cancer utilise compounds which inhibit DNAsynthesis. Such compounds are toxic to cells generally but their toxiceffect on rapidly dividing cells such as tumour cells can be beneficial.Alternative approaches to anti-proliferative agents which act bymechanisms other than the inhibition of DNA synthesis have the potentialto display enhanced selectivity of action.

In recent years it has been discovered that a cell may become cancerousby virtue of the transformation of a portion of its DNA into an oncogenei.e. a gene which, on activation, leads to the formation of malignanttumour cells (Bradshaw, Mutagenesis, 1986, 1, 91). Several suchoncogenes give rise to the production of peptides which are receptorsfor growth factors. The growth factor receptor complex subsequentlyleads to an increase in cell proliferation. It is known, for example,that several oncogenes encode tyrosine kinase enzymes and that certaingrowth factor receptors are also tyrosine kinase enzymes (Yarden et al.,Ann. Rev. Biochem., 1988, 57, 443. Larsen et al. Ann. Reports in Med.Chem. 1989, Chpt. 13).

Receptor tyrosine kinases are important in the transmission ofbiochemical signals which initiate cell replication. They are largeenzymes which span the cell membrane and possess an extracellularbinding domain for growth factors such as epidermal growth factor (EGF)and an intracellular portion which functions as a kinase tophosphorylate tyrosine amino acids in proteins and hence to influencecell proliferation. Various classes of receptor tyrosine kinases areknown (Wilks, Advances in Cancer Research, 1993. 60, 43-73) based onfamilies of growth factors which bind to different receptor tyrosinekinases. The classification includes Class I receptor tyrosine kinasescomprising the EGF family of receptor tyrosine kinases such as the EGF,TGFα, NEU, erbB, Xmrk, HER and let23 receptors, Class II receptortyrosine kinases comprising the insulin family of receptor tyrosinekinases such as the insulin, IGFI and insulin-related receptor (IRR)receptors and Class III receptor tyrosine kinases comprising theplatelet-derived growth factor (PDGF) family of receptor tyrosinekinases such as the PDGFα, PDGFβ and colony-stimulating factor I (CDF1)receptors. It is known that Class I kinases such as the EGF family ofreceptor tyrosine kinases are frequently present in common human cancerssuch as breast cancer (Sainsbury et al., Brit J. Cancer, 1988, 58, 458;Guerin et al, Oncogene Res., 1988, 3, 21 and Klijn et al., Breast CancerRes. Treat., 1994, 29, 73), non-small cell lung cancers (NSCLCs)including adenocarcinomas (Cerny et al., Brit. J. Cancer, 1986, 54, 265;Reubi et al., Int. J. Cancer, 1990, 45, 269; and Rusch et al., CancerResearch, 1993, 53, 2379) and squamous cell cancer of the lung (Hendleret al., Cancer Cells, 1989, 7, 347), bladder cancer (Neal et al.,Lancet, 1985, 366), oesophageal cancer (Mukaida et al., Cancer, 1991,68, 142), gastrointestinal cancer such as colon, rectal or stomachcancer (Bolen et al., Oncogene Res., 1987, 1, 149), cancer of theprostate (Visakorpi et al., Histochem. J., 1992, 24, 481), leukaemia(Konaka et al., Cell, 1984, 37, 1035) and ovarian, bronchial orpancreatic cancer (European Patent Specification No. 0400586). Asfurther human tumour tissues are tested for the EGF family of receptortyrosine kinases it is expected that their widespread prevalance will beestablished in further cancers such as thyroid and uterine cancer. It isalso known that EGF type tyrosine kinase activity is rarely detected innormal cells whereas it is more frequently detectable in malignant cells(Hunter, Cell, 1987, 50, 823). It has been shown more recently (W JGullick, Brit. Med. Bull., 1991, 47, 87) that EGF receptors whichpossess tyrosine kinase activity are overexpressed in many human cancerssuch as brain, lung squamous cell, bladder, gastric, breast, head andneck, oesophageal, gynaecological and thyroid tumours.

Accordingly it has been recognised that an inhibitor of receptortyrosine kinases should be of value as a selective inhibitor of thegrowth of mammalian cancer cells (Yaish et al. Science, 1988, 242, 933).Support for this view is provided by the demonstration that erbstatin,an EGF receptor tyrosine kinase inhibitor, specifically attenuates thegrowth in athymic nude mice of a transplanted human mammary carcinomawhich expresses EGF receptor tyrosine kinase but is without effect onthe growth of another carcinoma which does not express EGF receptortyrosine kinase (Toi et al., Eur. J. Cancer Clin. Oncol., 1990, 26,722). Various derivatives of styrene are also stated to possess tyrosinekinase inhibitory properties (European Patent Application Nos. 0211363,0304493 and 0322738) and to be of use as anti-tumour agents. The in vivoinhibitory effect of two such styrene derivatives which are EGF receptortyrosine kinase inhibitors has been demonstrated against the growth ofhuman squamous cell carcinoma inoculated into nude mice (Yoneda et al.,Cancer Research, 1991, 51, 4430). Various known tyrosine kinaseinhibitors are disclosed in a more recent review by T R Burke Jr. (Drugsof the Future, 1992, 17, 119).

It is known from European Patent Applications Nos. 0520722, 0566226 and0635498 that certain quinazoline derivatives which bear an anilinosubstituent at the 4-position possess receptor tyrosine kinaseinhibitory activity. It is further known from European PatentApplication No. 0602851 that certain quinazoline derivatives which beara heteroarylamino substituent at the 4-position also possess receptortyrosine kinase inhibitory activity.

It is further known from International Patent Application WO 92/20642that certain aryl and heteroaryl compounds inhibit EGF and/or PDGFreceptor tyrosine kinase. There is the disclosure of certain quinazolinederivatives therein but no mention is made of 4-anilinoquinazolinederivatives.

The in vitro anti-proliferative effect of a 4-anilinoquinazolinederivative has been disclosed by Fry et al., Science, 1994, 265, 1093.It was stated that the compound4-(3'-bromoanilino)-6,7-dimethoxyquinazoline was a highly potentinhibitor of EGF receptor tyrosine kinase.

The in vivo inhibitory effect of a 4,5-dianilinophthalimide derivativewhich is an inhibitor of the EGF family of receptor tyrosine kinases hasbeen demonstrated against the growth in BALB/c nude mice of a humanepidermnoid carcinoma A-431 or of a human ovarian carcinoma SKOV-3(Buchdunger et al., Proc. Nat. Acad. Sci., 1994, 91, 2334).

It is further known from European Patent Application No. 0635507 thatcertain tricyclic compounds which comprise a 5- or 6-membered ring fusedto the benzo-ring of a quinazoline possess receptor tyrosine kinaseinhibitory activity. It is also known from European Patent ApplicationNo. 0635498 that certain quinazoline derivatives which carry an aminogroup at the 6-position and a halogeno group at the 7-position possessreceptor tyrosine kinase inhibitory activity.

Accordingly it has been indicated that Class I receptor tyrosine kinaseinhibitors will prove to be useful in the treatment of a variety ofhuman cancers.

It is also expected that inhibitors of BGF type receptor tyrosinekinases will be useful in the treatment of other diseases of excessivecellular proliferation such as psoriasis (where TGFα is believed to bethe most important growth factor) and benign prostatic hypertrophy(BPH).

There is no disclosure in these documents of quinazoline derivativeswhich bear at the 4-position an anilino substituent and which also bearan aminoalkylamino substituent at the 6-position. We have now found thatsuch compounds possess potent in vivo anti-proliferative propertieswhich are believed to arise from their Class I receptor tyrosine kinaseinhibitory activity.

BRIEF SUMMARY OF THE INVENTION

According to the present invention there is provided a quinazolinederivative of the formula I ##STR1## wherein n is 1, 2 or 3 and each R²is independently halogeno; and R¹ isamino-(2-4C)alkylamino,(1-4C)alkylamino-(2-4C)alkylamino,di-[(1-4C)alkyl]amino-(2-4C)alkylamino,pyrrolidin-1-yl-(2-4C)alkylamino. piperidino-(2-4C)alkylamino,morpholino-(2-4C)alkylamino, piperazin-1-yl-(2-4C)alkylamino or4-(1-4C)alkylpiperazin-1-yl-(2-4C)alkylamino; or apharmaceutically-acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

In this specification the term "alkyl" includes both straight andbranched chain alkyl groups but references to individual alkyl groupssuch as "propyl" are specific to the straight chain version only. Forexample when R¹ is a di-[(1-4C)alkyl]amino-(2-4C)alkylamino group,suitable values for this generic radical include2-dimethylaminoethylamino, 3-dimethylaminopropylamino,2-dimethylaminopropylamino and 1-dimethylaminoprop-2-ylamino. Ananalogous convention applies to other generic terms.

Within the present invention it is to be understood that, insofar ascertain of the compounds of the formula I may exist in optically activeor racemic forms by virtue of one or more substituents containing anasymmetric carbon atom, the invention encompasses any such opticallyactive or racemic form which possesses anti-proliferative activity. Thesynthesis of optically active forms may be carried out by standardtechniques of organic chemistry well known in the art, for example bysynthesis from optically active starting materials or by resolution of aracemic form.

The quinazolines of the formula I are unsubstituted at the 2-, 5- and8-positions.

It is also to be understood that certain quinazoline derivatives of theformula I can exist in solvated as well as unsolvated forms such as, forexample, hydrated forms. It is to be understood that the inventionencompasses all such solvated forms which possess anti-proliferativeactivity.

Suitable values for the generic radicals referred to above include thoseset out below.

A suitable value for R² when it is halogeno is, for example fluoro,chloro, bromo or iodo.

Suitable values for each R¹ substituent which may be present on thequinazoline ring include, for example:

for amino-(2-4C)alkylamino: 2-aminoethylamino,3-aminopropylamino and4-aminobutylamino;

for (1-4C)alkylamino-(2-4C)alkylamino: 2-methylaminoethylamino,2-ethylaminoethylamino, 3-methylaminopropylamino,3-ethylaminopropylamino, 4-methylaminobutylamino and4-ethylaminobutylamino;

for di-[(1-4C)alkyl]amino-(2-4C)alkylamino: 2-dimethylaminoethylamino,2-(N-ethyl-N-methylamino)ethylamino. 2-diethylaminoethylamino,3-dimethylaminopropylamino, 3-diethylaminopropylamino and4-dimethylaminobutylamino;

for pyrrolidin-1-yl-(2-4C)alkylamino: 2-(pyrrolidin-1-yl)ethylamino,3-(pyrrolidin-1-yl)propylamino and 4-(pyrrolidin-1-yl)butylamino;

for piperidino-(2-4C)alkylamino: 2-piperidinoethylamino,3-piperidinopropylamino and 4-piperidinobutylamino;

for morpholino-(2-4C )alkylamino: 2-morpholinoethylamino,3-morpholinopropylamino and 4-morpholinobutylamino;

for piperazin-1-yl-(2-4C)alkylamino: 2-(piperazin-1-yl)ethylamino,3-(piperazin-1-yl)propylamino and 4-(piperazin-1-yl)butylamino;

for 4-(1-4C)alklipiperazin-1-yl-(2-4C)alkylamino:2-(4-methylpiperazin1-yl)ethylamino, 2-(4-ethylpiperazin-1-yl)ethylamino, 3-(4-methylpiperazin-1-yl)propylamino and4-(4-methylpiperazin-1-yl)butylamino.

A suitable pharmaceutically-acceptable salt of a quinazoline derivativeof the invention is, for example, an acid-addition salt of a quinazolinederivative of the invention which is sufficiently basic, for example, amono- or di-acid-addition salt with, for example, an inorganic ororganic acid, for example hydrochloric, hydrobromic, sulphuric,phosphoric, trifluoroacetic, citric, maleic, tartaric, fumaric,methanesulphonic or 4-toluenesulphonic acid.

Particular novel compounds of the invention include, for example,quinazoline derivatives of the formula I, or pharmaceutically-acceptablesalts thereof, wherein:

(a) n is 1 or 2 and each R² is independently fluoro, chloro or bromo:and R¹ has any of the meanings defined hereinbefore or in this sectionrelating to particular novel compounds of the invention;

(b) (R²)_(n) is 3'-fluoro, 3'-chloro, 3'-bromo, 2',4'-difluoro,2',4'-dichloro, 3',4'-difluoro, 3',4'-dichloro, 3'-fluoro-4'-chloro or3'-chloro-4'-fluoro; and R¹ has any of the meanings defined hereinbeforeor in this section relating to particular novel compounds of theinvention;

(c) R¹ is 2-aminoethylamino, 3-aminopropylamino,2-methylaminoethylamino, 3-methylaminopropylamino,2-dimethylaminoethylamino, 3-dimethylaminopropylamino,2-(pyrrolidin-1-yl)ethylamino,3-(pyrrolidin-1-yl)propylamino,2-piperidinoethylamino,3-piperidinopropylamino,2-morpholinoethylamino,3-morpholinopropylamino,2-(piperazin-1-yl)ethylamino,3-(piperazin-1-yl)propylanino,2-(4-methylpiperazin-1-yl)ethylamino or3-(4-methylpiperazin-1-yl)propylamino; and n and R² have any of themeanings defined hereinbefore or in this section relating to particularnovel compounds of the invention.

A preferred compound of the invention is a quinazoline derivative of theformula I wherein (R²)_(n) is 3'-chloro,3'-bromo, 3',4'-difluoro or3'-chloro-4'-fluoro; and R¹ is2-methylaminoethylamino,3-methylaminopropylamino,2-dimethylaminoethylamino,3-dimethylaminopropylamino,2-(pyrrolidin-1-yl)ethylamino,3-(pyrrolidin-1-yl)propylamino,2-piperidinoethylamino,3-piperidinopropylamino,2-morpholinoethylamino or3-morpholinopropylamino. or a pharmaceutically-acceptable acid-additionsalt thereof.

A further preferred compound of the invention is a quinazolinederivative of the formula I

wherein (R²)_(n) is 3',4 '-difluoro or 3'-chloro-4'-fluoro; and R¹ is3-dimethylaminopropylamino,3-(pyrrolidin-1-yl)propylamino,3-piperidinopropylamino or 3-morpholinopropylamino;

or a pharmaceutically-acceptable acid-addition salt thereof.

A quinazoline derivative of the invention, or apharmaceutically-acceptable salt thereof, may be prepared by any processknown to be applicable to the preparation of chemically-relatedcompounds. Suitable processes include, for example, those illustrated inEuropean Patent Applications Nos. 0520722, 0566226, 0602851, 0635498 and0635507. Such processes, when used to prepare a quinazoline derivativeof the invention, or a pharmaceutically-acceptable salt thereof, areprovided as a further feature of the invention and are illustrated bythe following representative Examples. Necessary starting materials maybe obtained by standard procedures of organic chemistry. The preparationof such starting materials is described within the accompanyingnon-limiting Examples. Alternatively necessary starting materials areobtainable by analogous procedures to those illustrated which are withinthe ordinary skill of an organic chemist.

(a) The reaction, conveniently in the presence of a suitable base, of aquinazoline of the formula II ##STR2## wherein Z is a displaceablegroup, with an aniline of the formula III ##STR3##

A suitable displaceable group Z is, for example, a halogeno, alkoxy,aryloxy or sulphonyloxy group, for example a chloro, bromo, methoxy,phenoxy, methanesulphonyloxy or toluene-4-sulphonyloxy group.

A suitable base is, for example, an organic amine base such as, forexample, pyridine, 2,6-lutidine, collidine, 4-dimethylaminopyridine,triethylamine, morpholine, N-methylmorpholine ordiazabicyclo[5.4.0]undec-7-ene, or, for example, an alkali or alkalineearth metal carbonate or hydroxide, for example sodium carbonate,potassium carbonate, calcium carbonate, sodium hydroxide or potassiumhydroxide. Alternatively a suitable base is, for example, an alkalimetal or alkaline earth metal amide, for example sodium amide or sodiumbis(trimethylsilyl)amide.

The reaction is preferably carried out in the presence of a suitableinert solvent or diluent, for example an alkanol or ester such asmethanol, ethanol, isopropanol or ethyl acetate, a halogenated solventsuch as methylene chloride, chloroform or carbon tetrachloride, an ethersuch as tetrahydrofuran or 1,4-dioxan, an aromatic solvent such astoluene, or a dipolar aprotic solvent such as N,N-dimethylformamide,N,N-dimethylacetamide, N-methylpyrrolidin-2-one or dimethylsulphoxide.The reaction is conveniently carried out at a temperature in the range,for example, 10 to 150° C., preferably in the range 20 to 80° C.

The quinazoline derivative of the invention may be obtained from thisprocess in the form of the free base or alternatively it may be obtainedin the form of a salt with the acid of the formula H--Z wherein Z hasthe meaning defined hereinbefore. When it is desired to obtain the freebase from the salt, the salt may be treated with a suitable base asdefined hereinbefore using a conventional procedure.

(b) For the production of those compounds of the formula I wherein R¹ isan amino-substituted (2-4C)alkylamino group, the alkylation,conveniently in the presence of a suitable base as defined hereinbefore,of a quinazoline derivative of the formula I wherein R¹ is an aminogroup.

A suitable alkylating agent is, for example, any agent known in the artfor the alkylation of amino to amino-substituted alkylamino, for examplean amino-substituted alkyl halide, for example an amino-substituted(2-4C)alkyl chloride, bromide or iodide in the presence of a suitablebase as defined hereinbefore.

The reaction is preferably carried out in a suitable inert solvent ordiluent as defined hereinbefore and at a temperature in the range, forexample, 0 to 150° C., conveniently at or near 90° C.

(c) For the production of these compounds of the formula I wherein R¹ isan amino-substituted (2-4C)alkylamino group, the reaction, convenientlyin the presence of a suitable base as defined hereinbefore, of acompound of the formula I wherein R¹ is a hydroxy-(2-4C)alkylaminogroup, or a reactive derivative thereof, with an appropriate amine.

A suitable reactive derivative of a compound of the formula I wherein R¹is a hydroxy-(2-4C)alkylamino group is, for example, a halogeno- orsulphonyloxy-(2-4C)alkylamino group such as a bromo- ormethanesulphonyloxy-(2-4C)alkylamino group.

The reaction is preferably carried out in a suitable inert solvent ordiluent as defined hereinbefore and at a temperature in the range, forexample, 0 to 150° C., conveniently at or near 70° C.

(d) The reductive amination of an amino-substituted aldehyde with acompound of the formula I wherein R¹ is amino.

The reduction may be carried out by any of the many procedures known forsuch a transformation. For example a hydride reducing agent may be used,for example an alkali metal borohydride or cyanoborohydride such assodium borohydride or sodium cyanoborohydride, or an alkali metalaluminium hydride such as lithium aluminium hydride. The reduction maybe carried out in the presence of a suitable inert solvent or diluent,for example a (1-4C)alcohol such as methanol or ethanol when an alkalimetal borohydride or cyanoborohydride is employed, or an inert ethersuch as diethyl ether or tetrahydrofuran when an alkali metal aluminiumhydride is employed.

(e) The reduction of a compound of the formula I wherein R¹ is anamino-substituted alkanoamido group.

The reduction may be carried out by any of the many procedures known forsuch a transformation. For example a hydride reducing agent may be used,for example an alkali metal aluminium hydride such as lithium aluminiumhydride. The reaction may be carried out in the presence of a suitableinert solvent or diluent, for example in an inert ether such as diethylether or tetrahydrofuran.

The reaction is conveniently carried out at a temperature in the range,for example, -10 to 100° C. conveniently at or near 60° C.

When a pharmaceutically-acceptable salt of a quinazoline derivative ofthe invention is required, for example a mono- or di-acid-addition saltof a quinazoline derivative of the invention, it may be obtained, forexample, by reaction of said compound with, for example, a suitable acidusing a conventional procedure.

According to a further aspect of the present invention there is providedthe quinazoline derivative4-(3'-chloro-4'-fluoroanilino)-6-[3-(pyrrolidin-1-yl)propylamino]-quinazoline;

or a pharmaceutically-acceptable salt thereof.

It is also to be understood that this quinazoline derivative of theinvention can exist in solvated as well as unsolvated forms such as, forexample, hydrated forms. It is to be understood that the inventionencompasses all such solvated forms which possess anti-proliferativeactivity.

This quinazoline derivative of the invention, or apharmaceutically-acceptable salt thereof, may be prepared as follows.

(a) The reaction, conveniently in the presence of a suitable base asdefined hereinbefore, of a quinazoline of the formula IV ##STR4##wherein Z is a displaceable group as defined hereinbefore, with3'-chloro-4'-fluoroaniline.

The reaction is preferably carried out in the presence of a suitableinert solvent or diluent, for example an alkanol or ester such asmethanol, ethanol, isopropanol or ethyl acetate, a halogenated solventsuch as methylene chloride, chloroform or carbon tetrachloride, an ethersuch as tetrahydrofliran or 1,4-dioxan, an aromatic solvent such astoluene, or a dipolar aprotic solvent such as N,N-dimethylformamide,N,N-dimethylacetamide, N-methylpyrrolidin-2-one or dimethylsulphoxide.The reaction is conveniently carried out at a temperature in the range,for example, 10 to 150° C., preferably in the range 20 to 80° C.

(b) The alkylation, conveniently in the presence of a suitable base asdefined hereinbefore, of4-(3'-chloro-4'-fluoroanilino)-6-aminoquinazoline with an alkylatingagent of the formula V ##STR5## wherein Z is a displaceable group asdefined hereinbefore.

The reaction is preferably carried out in a suitable inert solvent ordiluent as defined hereinbefore and at a temperature in the range, forexample, 0 to 150° C., conveniently at or near 90° C.

(c) The alkylation, conveniently in the presence of a suitable base asdefined hereinbefore of pyrrolidine with an alklylating agent of theformula VI ##STR6## wherein Z is a displaceable group as definedhereinbefore.

The reaction is preferably carried out in a suitable inert solvent ordiluent as defined hereinbefore and at a temperature in the range, forexample, 0 to 150° C., conveniently at or near 70° C.

(d) The reductive amination of 3-(pyrrolidin-1-yl)propionaldehyde with6-amino-4-(3'-chloro-4'-fluoroanilino)quinazoline.

The reduction may be carried out by any of the many procedures known forsuch a transformation. For example a hydride reducing agent may be used,for example an alkali metal borohydride or cyanoborohydride such assodium borohydride or sodium cyanoborohydride, or an alkali metalaluminium hydride such as lithium aluminium hydride. The reduction maybe carried out in the presence of a suitable inert solvent or diluent,for example a (1-4C)alcohol such as methanol or ethanol when an alkalimetal borohydride or cyanoborohydride is employed, or an inert ethersuch as diethyl ether or tetrahydrofuran when an alkali metal aluminiumhydride is employed. The reaction is conveniently carried out at atemperature in the range, for example, -10 to 100° C. conveniently at ornear ambient temperature.

As stated hereinbefore the quinazoline derivatives defined in thepresent invention possess anti-proliferative activity which is believedto arise from the Class I receptor tyrosine kinase inhibitory activityof the compounds. These properties may be assessed, for example, usingone or more of the procedures set out below:

(a) An in vitro assay which determines the ability of a test compound toinhibit the enzyme EGF receptor tyrosine kinase. Receptor tyrosinekinase was obtained in partially purified form from A-431 cells (derivedfrom human vulval carcinoma) by the procedures described below which arerelated to those described by Carpenter et al., J. Biol. Chem., 1979.254, 4884, Cohen et al., J. Biol. Chem., 1982, 257, 1523 and by Braun etal., J. Biol. Chem., 1984, 259, 2051.

A-431 cells were grown to confluence using Dulbecco's modified Eagle'smedium (DMEM) containing 5% fetal calf serum (FCS). The obtained cellswere homogenised in a hypotonic borate/EDTA buffer at pH 10.1. Thehomogenate was centrifuged at 400 g for 10 minutes at 0-4° C. Thesupernatant was centrifuged at 25,000 g for 30 minutes at 0-4° C. Thepelleted material was suspended in 30 mM Hepes buffer at pH 7.4containing 5% glycerol, 4 mM benzamidine and 1% Triton X-100, stirredfor 1 hour at 0-4° C. and recentrifuged at 100,000 g for 1 hour at 0-4°C. The supernatant, containing solubilised receptor tyrosine kinase, wasstored in liquid nitrogen.

For test purposes 40 μl of the enzyme solution so obtained was added toa mixture of 400 μl of a mixture of 150 mM Hepes buffer at pH 7.4. 500μM sodium orthovanadate, 0.1% Triton X-100, 10% glycerol, 200 μl water,80 μl of 25 mM DTT and 80 μl of a mixture of 12.5 mM manganese chloride,125 mM magnesium chloride and distilled water. There was thus obtainedthe test enzyme solution.

Each test compound was dissolved in dimethylsulphoxide (DMSO) to give a50 mM solution which was diluted with 40 mM Hepes buffer containing 0.1%Triton X-100, 10% glycerol and 10% DMSO to give a 500 μM solution. Equalvolumes of this solution and a solution of epidermal growth factor (EGF;20 μg/ml) were mixed.

[γ-³² P]ATP (3000 Ci/mM, 250 μCi) was diluted to a volume of 2 ml by theaddition of a solution of ATP (100 μM) in distilled water. An equalvolume of a 4 mg/ml solution of the peptideArg-Arg-Leu-Ile-Glu-Asp-Ala-Glu-Tyr-Ala-Ala-Arg-Gly in a mixture of 40mM Hepes buffer at pH 7.4, 0.1% Triton X-100 and 10% glycerol was added.

The test compound/EGF mixture solution (5 μl) was added to the testenzyme solution (10 μl) and the mixture was incubated at 0-4° C. for 30minutes. The ATP/peptide mixture (10 μl) was added and the mixture wasincubated at 25° C. for 10 minutes. The phosphorylation reaction wasterminated by the addition of 5% trichloroacetic acid (40 μl) and bovineserum albumin (BSA, 1 mg/ml, 5 μl). The mixture was allowed to stand at4° C. for 30 minutes and then centrifuged. An aliquot (40 μl) of thesupernatant was placed onto a strip of Whatman p 81 phosphocellulosepaper. The strip was washed in 75 mM phosphoric acid (4×10 ml) andblotted dry. Radioactivity present in the filter paper was measuredusing a liquid scintillation counter (Sequence A). The reaction sequencewas repeated in the absence of the EGF (Sequence B) and again in theabsence of the test compound (Sequence C).

Receptor tyrosine kinase inhibition was calculated as follows: ##EQU1##

The extent of inhibition was then determined at a range ofconcentrations of test compound to give an IC₅₀ value.

(b) An in vitro assay which determines the ability of a test compound toinhibit the EGF-stimulated growth of the human naso-pharyngeal cancercell line KB.

KB cells were seeded into wells at a density of 1×10⁴ -1.5×10⁴ cells perwell and grown for 24 hours in DMEM supplemented with 5% FCS(charcoal-stripped). Cell growth was determined after incubation for 3days by the extent of metabolism of MTT tetrazolium dye to furnish abluish colour. Cell growth was then determined in the presence of EGF(10 ng/ml) or in the presence of EGF (10 ng/ml) and a test compound at arange of concentrations. An ICSO value could then be calculated.

(c) An in-vivo assay in a group of athymic nude mice (strain ONU:Alpk)which determines the ability of a test compound (usually administeredorally as a ball-milled suspension in 0.5% polysorbate) to inhibit thegrowth of xenografts of the human vulval epidermoid carcinoma cell lineA-431.

A-431 cells were maintained in culture in DMEM supplemented with 5% FCSand 2 mM glutamine. Freshly cultured cells were harvested bytrypsinization and injected subcutaneously (10 million cells/0.1ml/mouse) into both flanks of a number of donor nude mice. Whensufficient tumour material was available (after approximately 9 to 14days), fragments of tumour tissue were transplanted in the flanks ofrecipient nude mice (test day 0). Generally, on the seventh day aftertransplantation (test day 7) groups of 7 to 10 mice with similar-sizedtumours were selected and dosing of the test compound was commenced.Once daily dosing of test compound was continued for a total of 13 days(test days 7 to 19 inclusive). In some studies the dosing of the testcompound was continued beyond test day 19, for example to test day 26.In each case, on the following test day the animals were killed and thefinal tumour volume was calculated from measurements of the length andwidth of the tumours. Results were calculated as a percentage inhibitionof tumour volume relative to untreated controls.

Although the pharmacological properties of the compounds of the formulaI vary with structural change as expected, in general activity possessedby compounds of the formula I may be demonstrated at the followingconcentrations or doses in one or more of the above tests (a), (b) and(c):

Test (a): IC₅₀ in the range, for example, 0.01-1 μM:

Test (b): lC₅₀ in the range, for example 0.05-1 μM:

Test (c): 20 to 90% inhibition of tumour volume from a daily dose in therange, for example, 12.5 to 200 mg/kg.

The compound of Example 1 of the invention possesses activity atapproximately the following concentrations or doses in tests (a), (b)and (c):

Test (a): IC₅₀ 0.076 μM;

Test (b): IC₅₀ 0.06 μM;

Test (c): greater than 50% inhibition of tumour volume from a daily doseof 50 mg/kg.

According to a further aspect of the invention there is provided apharmaceutical composition which comprises a quinazoline derivative ofthe formula I, or a pharmaceutically-acceptable salt thereof, inassociation with a pharmaceutically-acceptable diluent or carrier.

The composition may be in a form suitable for oral administration, forexample as a tablet or capsule, for parenteral injection (includingintraveous, subcutaneous, intramuscular, intravascular or infusion) as asterile solution, suspension or emulsion, for topical administration asan ointment or cream or for rectal administration as a suppository.

In general the above compositions may be prepared in a conventionalmanner using conventional excipients.

The quinazoline derivative will normally be administered to awarm-blooded animal at a unit dose within the range 5-10000 mg persquare meter body area of the animal, i.e. approximately 0.1-200 mg/kg.and this normally provides a therapeutically-effective dose. A unit doseform such as a tablet or capsule will usually contain, for example 1-250mg of active ingredient. Preferably a daily dose in the range of 1-100mg/kg is employed. However the daily dose will necessarily be varieddepending upon the host treated, the particular route of administration,and the severity of the illness being treated. Accordingly the optimumdosage may be determined by the practitioner who is treating anyparticular patient.

According to a further aspect of the present invention there is provideda quinazoline derivative of the formula I, or apharmaceutically-acceptable salt thereof, for use in a method oftreatment of the human or animal body by therapy.

We have now found that the compounds of the present invention possessanti-proliferative properties such as anti-cancer properties which arebelieved to arise from their Class I receptor tyrosine kinase inhibitoryactivity. Accordingly the compounds of the present invention areexpected to be useful in the treatment of diseases or medical conditionsmediated alone or in part by Class I receptor tyrosine kinases, i.e. thecompounds may be used to produce a Class I receptor tyrosine kinaseinhibitory effect in a warm-blooded animal in need of such treatment.Thus the compounds of the present invention provide a method fortreating the proliferation of malignant cells characterised byinhibition of Class I receptor tyrosine kinases, i.e. the compounds maybe used to produce an anti-proliferative effect mediated alone or inpart by the inhibition of Class I receptor tyrosine kinase. Accordinglythe compounds of the present invention are expected to be useful in thetreatment of psoriasis and/or cancer by providing an anti-proliferativeeffect, particularly in the treatment of Class I receptor tyrosinekinase sensitive cancers such as cancers of the breast, lung, colon,rectum, stomach, prostate, bladder, pancreas and ovary.

Thus according to this aspect of the invention there is provided the useof a quinazoline derivative of the formula I, or apharmaceutically-acceptable salt thereof, in the manufacture of amedicament for use in the production of an anti-proliferative effect ina warm-blooded animal such as man.

According to a further feature of this aspect of the invention there isprovided a method for producing an anti-proliferative effect in awarm-blooded animal, such as man, in need of such treatment whichcomprises administering to said animal an effective amount of aquinazoline derivative as defined immediately above.

As stated above the size of the dose required for the therapeutic orprophylactic treatment of a particular proliferative disease willnecessarily be varied depending on the host treated, the route ofadministration and the severity of the illness being treated. A unitdose in the range, for example, 1-200 mg/kg, preferably 1-100 mg/kg isenvisaged.

The anti-proliferative treatment defined hereinbefore may be applied asa sole therapy or may involve, in addition to the quinazoline derivativeof the invention, one or more other anti-tumour substances, for examplethose selected from, for example, mitotic inhibitors, for examplevinblastine, vindesine and vinorelbine; tubulin disassembly inhibitorssuch as taxol; alkylating agents, for example cis-platin, carboplatinand cyclophosphamide; antimetabolites, for example 5-fluorouracil,tegafur, methotrexate, cytosine arabinoside and hydroxyurea, or, forexample, one of the preferred antimetabolites disclosed in EuropeanPatent Application No. 239362 such asN-{5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl}-L-glutamicacid; intercalating antibiotics, for example adriamycin, mitomycin andbleomycin; enzymes, for example asparaginase; topoisomerase inhibitors,for example etoposide and camptothecin; biological response modifiers,for example interferon; and anti-hormones, for example antioestrogenssuch as tamoxifen or, for example antiandrocens such as4'-cyano-3-(4-fluorophenysulphonyl)-2-hydroxy-2-methyl-3'-(trifluoromethyl)-propionanilide.Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of thetreatment. According to this aspect of the invention there is provided apharmaceutical product comprising a quinazoline derivative of theformula I as defined hereinbefore and an additional anti-tumoursubstance as defined hereinbefore for the conjoint treatment of cancer.

As stated above the quinazoline derivative defined in the presentinvention is an effective anti-cancer agent, which property is believedto arise from its Class I receptor tyrosine kinase inhibitoryproperties. Such a quinazoline derivative of the invention is expectedto possess a wide range of anti-cancer properties as Class I receptortyrosine kinases have been implicated in many common human cancers suchas leukaemia and breast, lung, colon, rectal, stomach, prostate,bladder, pancreas and ovarian cancer. Thus it is expected that aquinazoline derivative of the invention will possess anti-canceractivity against these cancers. It is in addition expected that aquinazoline derivative of the present invention will possess activityagainst a range of leukaemias, lymphoid malignancies and solid tumourssuch as carcinomas and sarcomas in tissues such as the liver, kidney,prostate and pancreas.

It is further expected that a quinazoline derivative of the inventionwill possess activity against other diseases involving excessivecellular proliferation such as psoriasis. benign prostatic hypertrophy(BPH) and atherosclerosis.

It is also to be expected that a quinazoline derivative of the inventionwill be useful in the treatment of additional disorders of cellulargrowth in which aberrant cell signalling by way of receptor tyrosinekinase enzymes, including as yet unidentified receptor tyrosine kinaseenzymes, are involved. Such disorders include for example, inflammation,angiogenesis, vascular restenosis, immunological disorders,pancreatitis, kidney, disease and blastocyte maturation andimplantation.

The invention will now be illustrated in the following non-limitingExamples in which, unless otherwise stated:

(i) evaporations were carried out by rotary evaporation in vacuo andwork-up procedures were carried out after removal of residual solidssuch as drying agents by filtration, unless otherwise stated magnesiumsulphate was used as a drying agent for organic solutions;

(ii) operations were carried out at ambient temperature, that is in therange 18-25° C. and under an atmosphere of an inert gas such as argon;

(iii) column chromatography (by the flash procedure) and medium pressureliquid chromatography (MPLC) were performed on Merck Kieselgel silica(Art. 9385) or Merck Lichroprep RP-18 (Art. 9303) reversed-phase silicaobtained from E. Merck, Darmstadt, Germany;

(iv) yields are given for illustration only and are not necessarily themaximum attainable;

(v) melting points were determined using a Mettler SP62 automaticmelting point apparatus, an oil-bath apparatus or a Koffler hot plateapparatus.

(vi) the structure of the compound of the invention was confirmed bynuclear (generally proton) magnetic resonance (NMR) and mass spectraltechniques; proton magnetic resonance chemical shift values weremeasured on the delta scale and peak multiplicities are shown asfollows: s, singlet; d, doublet; t, triplet; m, multiplet, unlessotherwise stated end-products of the invention were dissolved in CD₃SOCD₃ for the determination of NMR values;

(vii) intermediates were not generally fully characterised and puritywas assessed by thin layer chromatography (TLC), infrared (IR) or NMRanalysis.

(viii) the following abbreviations have been used:

DMF N,N-dimethylformamide:

THF tetrahydrofuran;

DMA N,N-dimethylacetamide.

EXAMPLE 1

A mixture of 3-(pyrrolidin-1-yl)propionaldehyde dimethyl acetal (2.4 g)and water (12 ml) was acidified to pH1 by the addition of concentratedhydrochloric acid and the mixture was stirred and heated to 40° C. for90 minutes. The mixture was then reduced in acidity to pH6 by theaddition of 1M aqueous sodium bicarbonate solution. The resultantmixture was added to a mixture of6-amino-4-(3'-chloro-4'-fluoroanilino)quinazoline (1.64 g) and sodiumcyanoborohydride (0.41 g). The mixture was acidified to pH4 by theaddition of glacial acetic acid and stirred at ambient temperature for16 hours. The mixture was evaporated and the residue was partitionedbetween methylene chloride and water. The organic phase was washed withwater, dried (MgSO₄) and evaporated. The residue was purified by columnchromatography using increasingly polar mixtures of methylene chlorideand methanol [each mixture also containing 0.1% of a concentrated (30%weight/volume) aqueous ammonium hydroxide solution]. The material soobtained was triturated under diethyl ether. There was thus obtained4-(3'-chloro-4'-fluoroanilino)-6-[3-(pyrrolidin-1-yl)propylamino]quinazoline (1.1 g, 48%), m.p. 155-157° C.;

NMR Spectrum: (CD₃ SOCD₃ +CD₃ CO₂ D) 1.94-2.22 (m, 6H), 3.22-3.43 (m,8H), 7.27 (D, 1H), 7.32 (m, 1H), 7.38 (t, 1H), 7.62 (d, 1H), 7.87 (m,1H), 8.18 (m, 1H) 8.42 (s, 1H);

Elemental Analysis: Found C, 60.7; H, 5.8; N, 16.7;

C₂₁ H₂₃ CIFN₅ 1H₂ O requires C, 60.4; H, 6.0; N. 16.8%.

The 6-amino-4-(3'-chloro-4'-fluoroanilino)quinazoline used as a startingmaterial was obtained as follows:

3'-Chloro-4'-fluoroaniline (3.6 g) was added to a stirred mixture of4-chloro-6-nitroquinazoline (European Patent Application No. 0566226.Example 8 thereof; 5 g); THF (10 ml) and DMF (10 ml). The resultantmixture was stirred at ambient temperature for 5 hours. The precipitatewas isolated and partitioned between water and a 9:1 mixture ofmethylene chloride and methanol. The aqueous phase was neutralised bythe addition of a saturated aqueous sodium bicarbonate solution andre-extracted with methylene chloride. The organic phases were combinedand evaporated. The residue was triturated under a 9:1 mixture ofethanol and water. The resultant solid was isolated and dried. There wasthus obtained 4-(3'-chloro-4'-fluoroanilino)-6-nitroquinazoline (2.5 g).

A mixture of a portion (2.3 g) of the material so obtained, 10%palladium-on-carbon catalyst (0.4 g), ethanol (25 ml) and DMF (25 ml)was stirred under an atmosphere of hydrogen for 2 hours. The mixture wasfiltered and the filtrate was evaporated. The residue was trituratedunder a 4:1 mixture of ethanol and water. The resultant solid wasisolated and dried. There was thus obtained6-amino-4-(3'-chloro-4'-fluoroanilino)quinazoline (0.35 g, 17%);

NMR Spectrum: 5.6 (broad s, 2H), 7.27 (m, 1H), 7.32 (s, 1H), 7.41 (t,1H), 7.55 (d, 1H), 7.8 (m, 1H), 8.19 (m, 1H), 8.38 (s, 1H), 9.53 (broads, 1H);

Elemental Analysis: Found C, 58.1; H, 3.6; N, 19.0;

C₁₄ H₁₀ CIFN₄ requires C, 58.2; H. 3.5; N, 19.4%.

The 3-(pyrrolidin-1-yl)propionaldehyde dimethyl acetal used as astarting material was obtained as follows:

Pyrrolidine (7.8 g) was added to a stirred mixture of3-bromopropionaldehyde dimethyl acetal (10 g), potassium carbonate (22.6g) and toluene (150 ml). The mixture was stirred at ambient temperaturefor 16 hours. The mixture was filtered and the filtrate was evaporated.There was thus obtained 3-(pyrrolidin-1-yl)propionaldehyde dimethylacetal (8.9 g) which was used without further purification;

NMR Spectrum: 1.65 (m 6H), 2.40 (m, 6H), 3.2 (s, 6H), 4.4 (t, 1H).

EXAMPLE 2

Using an analogous procedure to that described in Example 1,6-amino-4-(3'-chloro-4'-fluoroanilino)quinazoline was reacted with3-morpholinopropionaldehyde dimethyl acetal to give4-(3'-chloro-4'-fluoroanilino)-6-(3-morpholinopropylarnino)quinazolinein 84% yield;

NMR Spectrum: 1.63 (m, 2H), 2.48 (m, 6H), 3.63 (t, 4H), 6.23 (t, 1H),7.27 (d, 1H), 7.32 (m, 1H), 7.43 (t, 1H), 7.62 (d, 1H), 7.84 (m, 1H),8.15 (m, 1H), 8.37 (s, 1H), 9.42 (broad s, 1H);

Elemental Analysis: C, 59.4; H. 5.9; N, 16.0;

C₂₁ H₂₃ CIFN₅ O 0.55H₂ O requires C, 59.2; H, 5.7; N, 16.4%.

The 3-morpholinopropionaldehyde dimethyl acetal used as a startingmaterial was obtained by the reaction of morpholine and3-bromopropionaldehyde dimethyl acetal using an analgous procedure tothat described in the last portion of Example 1.

EXAMPLE 3

A 1M solution of lithium aluminium hydride (35 ml) was added to asolution of4-(3',4'-difluoroanilino)-6-(2-methylaminoacetamido)quinazoline (3.3 g)in THF (200 ml) and the resultant mixture was stirred and heated toreflux for 2 hours. The mixture was cooled to ambient temperature andglacial acetic acid was added to destroy the excess of reducing agent.The mixture was evaporated and the residue was partitioned betweenmethylene chloride and a dilute aqueous ammonium hydroxide solution. Theorganic phase was dried and evaporated and the residue was purified bycolumn chromatography using increasingly polar mixtures of methylenechloride, methanol and a dilute (2%) aqueous ammonium hydroxidesolution. The material so obtained was triturated under diethyl ether.There was thus obtained4-(3',4'-difluoroanilino)-6-(2-methylaminoethylamino)quinazoline (0.61g), m.p. 146-148° C.;

NMR Spectrum: 2.5 (s, 3H), 2.85 (t, 2H), 3.3 (t, 2H), 6.15 (t, 1H), 7.2(d, 1H), 7.3(m, 1H), 7.4 (m, 1H), 7.55 (d, 1H), 7.65 (m, 1H), 8.05-8.2(m, 2H), 8.4 (s, 1H), 9.45 (broad d, 1H);

Elemental Analysis: Found C, 59.6; H, 5.3; N, 20.4;

C₁₇ H₁₇ F₂ N₅ 0.75H₂ O requires C, 59.6; H, 5.4; N, 20.4%.

The 4-(3',4'-difluoroanilino)-6-(2-methylaminoacetamido)quinazoline usedas a starting material was obtained as follows:

3',4'-difluoroaniline (27 g) was added to a stirred mixture of4-chloro-6-nitroquinazoline (44 g), triethylamine (29.2 ml) andmethylene chloride (400 ml) and the mixture was stirred at ambienttemperature for 16 hours. The precipitate was isolated, washed withmethylene chloride and acetone and dried. There was thus obtained4-(3',4'-difluoroanilino)-6-nitroquinazoline hydrochloride (61.3 g,82%).

A mixture of a portion (30 g) of the material so obtained, 10%palladium-on-carbon catalyst (3 g), ethanol (350 ml) and DMA (700 ml)was stirred under an atmosphere of hydrogen for 9 hours. The mixture wasfiltered and the filtrate was evaporated. The residual solid was washedin turn with DMA, with ethanol and with diethyl ether and dried. Therewas thus obtained 6-amino-4-(3',4'-difluoroanilino)quinazolinehydrochloride (12.8 g, 40%), m.p. 235-238° C.;

NMR Spectrum: 7.45 (m, 1H), 7.56 (m, 3H), 7.77 (d, 1H), 7.91 (m, 1H),8.72 (s, 1H), 11.1 (broad s, 1H);

Elemental Analysis: Found C, 54.5; H, 3.5; N, 18.1;

C₁₄ H₁₀ F₂ N₄ HCl requires C, 54.5; H, 3.6; N, 18.1%.

A concentrated aqueous solution of ammonium hydroxide (50 ml) was addedto a stirred suspension of 6-amino-4-(3',4'-difluoroanilino)quinazolinehydrochloride (5 g) in water (60 ml) and the mixture was stirred atambient temperature for 40 minutes. The mixture was filtered and thesolid so obtained was washed with water and dried. The material soobtained was dissolved in DMA (90 ml), 2-chloroacetyl chloride (1.29 ml)was added and the mixture was stirred at ambient temperature for 16hours. The mixture was evaporated and the residue was dried. There wasthus obtained6-(2-chloroacetamido)-4-(3',4'-difluoroanilino)-quinazolinehydrochloride (5.72 g, 91%). m.p.>250° C.;

NMR Spectrum: 4.43 (s, 2H), 7.54 (m, 2H), 7.97 (m, 1H), 7.99 (d, 1H),8.08 (m, 1H), 8.89 (s, 1H), 9.02 (d, 1H), 11.2 (broad s, 1H), 11.6(broad s, 1H);

Elemental Analysis: Found C, 49.4; H, 3.1; N, 14.3;

C₁₆ H₁₁ ClF₂ N₄ O HCl requires C, 49.9; H, 3.1; N, 14.5%.

Methylamine (33% solution in ethanol; 50 ml) was added to a stirredmixture of 6-(2-chloroacetamido)-4-(3',4'-difluoroanilino)quinazolinehydrochloride (3.3 g) and DMA (200 ml) and the resultant mixture wasstirred at ambient temperature for 2.5 hours. The mixture was evaporatedand the residue was triturated under an aqueous ammonium hydroxidesolution. The resultant precipitate was isolated, washed with water andwith diethyl ether and dried. There was thus obtained4-(3',4'-difluoroanilino)-6-(2-methylaminoacetamido)quinazoline (3.5 g);

NMR Spectrum: 2.4 (s, 3H), 3.35 (s, 2H), 7.4 (m, 1H), 7.6 (m, 1H), 7.8(d, 1H), 8.0-8.15 (m, 2H), 8.55 (s, 1H), 8.6 (d, 1H), 9.75 (broad s,1H);

Elemental Analysis: Found C. 56.2; H, 4.6; N, 19.1;

C₁₇ H₁₅ F₂ N₅ O H₂ O requires C, 56.5; H, 4.7; N, 19.4%.

EXAMPLE 4

Using an analogous procedure to that described in Example 1,4-(3',4'-difluoroanilino)-6-(2-morpholinoacetamido)quinazoline wasreduced with lithium aluminium hydride to give4-(3',4'-difluoroanilino)-6-(2-morpholinoethylamino)quinazoline in 58%yield;

NMR Spectrum: 2.48 (t, 4H), 2.64 (t, 2H), 3.32 (m, 2H), 3.62 (t, 4H),6.02 (t, 1H), 7.2 (d, 1H), 7.32 (m, 1H), 7.42 (m, 1H), 7.53 (d, 1H), 7.6(m, 1H), 8.08 (m, 1H), 8.38 (s, 1H), 9.4 (broad s, 1H);

Elemental Analysis: Found C, 62.6; H, 5.7; N, 17.7;

C₂₀ H₂₁ F₂ N₅ O 0.02CH₃ CO₂ H requires C, 62.2; H, 5.5; N, 18.1%.

The 4-(3',4'-difluoroanilino)-6-(2-morpholinoacetamido)quinazoline usedas a starting material was obtained as follows:

Morpholine (20 g) was added to a stirred solution of6-(2-chloroacetamido)-4-(3',4'-difluoroanilino)quinazoline hydrochloride(1.87 g) in DMA (50 ml) and the mixture was stirred at ambienttemperature for 40 hours. The mixture was evaporated and the residue waspurified by column chromatography using increasingly polar mixtures ofmethylene chloride and methanol as eluent. There was thus obtained4-(3',4'-difluoroanilino)-6-(2-morpholinoacetamido)quinazoline (1.78 g92%). m.p. 177-178° C.;

NMR Spectrum: 2.55 (m, 4H), 3.21 (s, 1H), 3.68 (t, 4H), 7.42 (m 1H),7.63 (m, 1H), 7.79 (d, 1H), 8.03 (m, 1H), 8.07 (m, 1H), 8.55 (s, 1H),8.62 (d, 1H), 9.83 (broad s, 1H), 9.98 (broad s, 1H);

Elemental Analysis: Found C, 59.7; H, 4.8; N, 17.0;

C₂₀ H₁₉ F₂ N₅ O₂ 0.2H₂ O requires C, 59.6; H, 4.85; N, 17.4%.

EXAMPLE 5

The following illustrate representative pharmaceutical dosage formscontaining the compound of formula I, or a pharmaceutically-acceptablesalt thereof (hereafter compound X), for therapeutic or prophylactic usein humans:

    ______________________________________                                                          mg/tablet                                                   ______________________________________                                        (a)  Tablet I                                                                      Compound X         100                                                        Lactose Ph. Eur    182.75                                                     Croscarmellose sodium                                                                            12.0                                                       Maize starch paste (5% w/v paste)                                                                2.25                                                       Magnesium stearate 3.0                                                   (b)  Tablet II                                                                     Compound X         50                                                         Lactose Ph. Eur    223.75                                                     Croscarmellose sodium                                                                            6.0                                                        Maize starch       15.0                                                       Polyvinylpyrrolidone                                                                             2.25                                                       Magnesium stearate 3.0                                                   (c)  Tablet III                                                                    Compound X         1.0                                                        Lactose Ph. Eur    93.25                                                      Croscarmellose sodium                                                                            4.0                                                        Maize starch paste (5% w/v paste)                                                                0.75                                                       Magnesium stearate 1.0                                                   ______________________________________                                        (d)  Capsule            mg/capsule                                            ______________________________________                                             Compound X         10                                                         Lactose Ph. Eur    488.5                                                      Magnesium stearate 1.5                                                   ______________________________________                                        (e)  Injection I        (50 mg/ml)                                            ______________________________________                                             Compound X          5.0% w/v                                                  1 M Sodium hydroxide solution                                                                    15.0% w/v                                                  0.1 M Hydrochloric acid                                                       (to adjust pH to 7.6)                                                         Polyethylene glycol 400                                                                           4.5% w/v                                                  Water for injection to 100%                                              ______________________________________                                        (f)  Injection II       (10 mg/ml)                                            ______________________________________                                             Compound X          1.0% w/v                                                  Sodium phosphate BP                                                                               3.6% w/v                                                  0.1 M Sodium hydroxide solution                                                                  15.0% w/v                                                  Water for injection to 100%                                              ______________________________________                                        (g)  Injection III      (1 mg/ml, buffered to pH 6)                           ______________________________________                                             Compound X          0.1% w/v                                                  Sodium phosphate BP                                                                              2.26% w/v                                                  Citric acid        0.38% w/v                                                  Polyethylene glycol 400                                                                           3.5% w/v                                                  Water for injection to 100%                                              ______________________________________                                    

Note

The above formulations may be obtained by conventional procedures wellknown in the pharmaceutical art. The tablets (a)-(c) may be entericcoated by conventional means, for example to provide a coating ofcellulose acetate phthalate.

I claim:
 1. A quinazole derivative of the formula I ##STR7## wherein nis 1, 2 or 3 and each R² is independently halogeno; and R¹ isamino-(2-4C)alkylamino, (1-4C)alkylamino-(2-4C)alkylamino,di-[(1-4C)alkyl]amino-(2-4C)alkylamino,pyrrolidin-1-yl-(2-4C)alkylamino, piperidino-(2-4C)alkylamino,morpholino-(2-4C)alkylamino, piperazin-1-yl-(2-4C)alkylamino or4-(1-4C)alkylpiperazin-1-yl-(2-4C)alkylamino;or apharmaceutically-acceptable salt thereof.
 2. A quinazoline derivative ofthe formula I as claimed in claim 1 wherein (R²)_(n) is3'-chloro,3'-bromo,3',4'-difluoro or 3'-chloro-4'-fluoro; and R¹ is2-methylaminoethylamino,3-methylaminopropylamino,2-dimethylaminoethylamino,3-dimethylaminopropylamino,2-(pyrrolidin-1-yl)ethylamino,3-(pyrrolidin-1-yl)propylamino,2-piperidinoethylamino,3-piperidinopropylamino,2-morpholinoethylamino or3-morpholinopropylamino;or a pharmaceutically-acceptable acid-additionsalt thereof.
 3. A quinazoline derivative of the formula I as claimed inclaim 1 wherein (R²)_(n) is 3',4'-difluoro or 3'-chloro-4'-fluoro; andR¹ is 3-dimethylaminopropylamino, 3-(pyrrolidin-1-yl)propylamino,3-piperidinopropylamino or 3-morpholinopropylamino;or apharmaceutically-acceptable acid-addition salt thereof.
 4. Thequinazoline derivative of the formula I as claimed in claim 1being:4-(3'-chloro-4'-fluoroanilino)-6-[3-(pyrrolidin-1-yl)propylamino]quinazoline;ora pharmaceutically-acceptable salt thereof.
 5. A process for thepreparation of a quinazoline derivative of the formula I, or apharmaceutically-acceptable salt thereof, as claimed in any one ofclaims 1 to 4 which comprises:(a) the reaction of a quinazoline of theformula II ##STR8## wherein Z is a displaceable group, with an anilineof the formula III ##STR9## (b) for the production of those compounds ofthe formula I wherein R¹ is an amino-substituted (2-4C)alkylamino group,the alkylation of a quinazoline derivative of the formula I wherein R¹is an amino group;and when a pharmaceutically-acceptable salt of aquinazoline derivative of the invention is required it may be obtainedby reaction of said compound with a suitable acid using a conventionalprocedure.
 6. A pharmaceutical composition which comprises a quinazolinederivative of the formula I, or a pharmaceutically-acceptable saltthereof, as claimed in any one of claims 1 to 4 in association with apharmaceutically-acceptable diluent or carrier.
 7. A method forproducing an anti-proliferative effect in a warm-blooded animal in needof such treatment which comprises administering to said animal aneffective amount of a quinazoline derivative of the formula I, or apharmaceutically-acceptable salt thereof, as claimed in any one ofclaims 1 to
 4. 8. A pharmaceutical product comprising a quinazolinederivative of the formula I, or a pharmaceutically-acceptable saltthereof, as claimed in any one of claims 1 to 4 and an additionalanti-tumour substance for the conjoint treatment of cancer.
 9. A methodfor producing a Class I receptor tyrosine kinase inhibitory effect in awarm-blooded animal in need of such treatment which comprisesadministering to said animal an effective amount of quinazolinederivative of the formula I, or a pharmaceutically-acceptable saltthereof, as claimed in any one of claims 1 to 4.